China Oncology ›› 2014, Vol. 24 ›› Issue (4): 258-265.doi: 10.3969/j.issn.1007-3969.2014.04.004

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Isolation, identification, and senescence examination of cervical cancer and normal cervical fibroblasts

CHEN Wei1, CHEN Ya-ping1, YANG Gong2,3   

  1. 1.Department of Gynecology and Obstetrics, the 5th People’s Hospital Affiliated to Fudan University; Department of Gynecology and Obstetrics, Shanghai Medical College, Fudan University, Shanghai 200240; 2.Central Laboratory, the 5th People’s Hospital Affiliated to Fudan University; Department of Gynecology and Obstetrics, Shanghai Medical College, Fudan University, Shanghai 200240; 3.Cancer Research Laboratory, Fudan University Shanghai Cancer Center; Depatment of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China
  • Online:2014-04-30 Published:2014-05-06
  • Contact: YANG Gong E-mail: yanggong@fudan.edu.cn

Abstract:

Background and purpose: Cervial cancer is most common gynecological malignancy. In recent years, the number of the young patients has obviously risen. It is now believed that almost all cases of cervical cancer are caused by exposure to a high-risk human papilloma virus (HPV). It has demonstrated that epithelial cancer can induce the senescence of normal stromal fibroblasts and senescent fibroblasts can promote tumor growth. This study by comparing the secretion of interleukin-6(IL-6) and vascular endothelial growth factor(VEGF) helped us to understand the mechanism of cervical cancer, and providesd more adequate theoretical and experimental basis for clinical early diagnosis and treatment of the disease. This study characterized normal fibroblasts (NFs) and cancer-associated fibroblasts (CAFs) to understand the mechanism how the chronic inflammation induced cervical cancer, and to provide novel theoretical basis for clinical diagnosis and treatment of the disease. Methods: NFs and CAFs were isolated and purified from normal and cervical cancer tissues. The expression levels of IL-6 and VEGF in cell culture medium were measured by ELISA and cellular senescence was detected by senescence-associated beta galactosidase (SA-β-gal) staining. The senescent marker p16 was analyzed by Western blot. Results: Our data showed that the expressions of both IL-6 and VEGF were higher in CAFs than in NFs (P<0.05), and that the blue staining of SA-β-gal and the expression of p16 and VEGF were stronger in CAFs than in NFs. Conclusion: Cervical epithelial cancer may be resulted from the senescence of stromal fibroblasts and HPV infection. Studying on cytokine-mediated stromal senescence may have a potential value for prevention, diagnosis, and treatment of cervical cancer.

Key words: Cervical cancer, Stromal fibroblast, Cellular senescence, Epithelial cancer cell